Upland Transformation And Offspring Analysis Of Different LEA Gene With Resilience

As an agricultural country,China has been conducting several of studies on the rice breeding.Recently,water is increasingly scarce and the yield and quality of rice was seriously affected.Therefore,studies of rice which can use less water and has drought-resistance become particularly important.Upland is easier to survive under dry conditions than rice.But there are some shortcomings in the cultivation of upland.Many bad factors such as weeds,soil salinization affect the yield and quality of upland to some extent.Therefore,improving the traits of upland is also of great value.In this study,“Kunhan 1”has been choosed as the material of experiment.Three different kinds of gene named 8981、DEHY、8704,which from family LEA（Late Embryogenesis Abundant Proteins）were transformed to“Kunhan 1”by Agrobacterium transformation.Different gene LEA types of transgenic upland had been achieved in this experiment and the results of the three types of genes had been compared and analyzed.The positive identification of T₁transgenic plants on DNA level and the preliminary screening of T₂transgenic lines had been conducted.In the process of callus induction,the quality and quantity of callus had been increased by shoots cut.There are some differences in the rates of green dots and differentiations,which is known by comparing the results of the three gene types.Followed by the 8981gene,the rates of green dots and differentiations of Transgenic Plants with DEHY gene is the highest,and the 8704 gene is the lowest one.In the process of homozygotes screening by herbicide,the optimum concentration of screening had been determined as 0.5mg/m L.Lines suspected homozygous had been selected from positive plants of generation 2 preliminary.During the analysis of Southern hybridization,the optimum sample volume and temperature of washing stringent had been determined as 15μg and 68℃by making adjustments to the hybrid system.Finally,homozygous lines which have just a single copy of transgenic gene LEA and can stably inherited had been screened.